Lei Ye

Pathogens present significant risks to public health and typically require detection in a hospital or clinic setting. The advancement of lateral flow assay (LFA) cassettes has made detection accessible to all, allowing individuals to perform tests from nearly any location. The extensive use of antigen rapid test (ART) kits for self-screening of COVID has reinforced the critical role of LFAs in accelerating diagnostic processes and subsequent disease prevention measures. This study delves into the realm of lateral flow assays, encompassing their fundamental design principles, interpretation of results, and the cutting-edge research focused on enhancing reporter materials and signal amplification techniques. Additionally, it emphasizes recent progress in employing LFAs for the effective identification of a diverse array of pathogenic microorganisms. This comprehensive overview has the potential to serve as a valuable reference for researchers in related disciplines.

Pathogens present significant risks to public health and typically require detection in a hospital or clinic setting. The advancement of lateral flow assay (LFA) cassettes has made detection accessible to all, allowing individuals to perform tests from nearly any location. The extensive use of antigen rapid test (ART) kits for self-screening of Covid-19 has reinforced the critical role of LFAs in accelerating diagnostic processes and subsequent disease prevention measures. This study delves into the realm of lateral flow assays, encompassing their fundamental design principles, interpretation of results, and the cutting-edge research focused on enhancing reporter materials and signal amplification techniques. Additionally, it emphasizes recent progress in employing LFAs for the effective identification of a diverse array of pathogenic microorganisms. This comprehensive overview has the potential to serve as a valuable reference for researchers in related disciplines.

Peripheral arterial disease (PAD) is estimated to affect at least 10 million people in the United States and over 200 million individuals worldwide. The clinical manifestations of PAD range from impaired mobility and pain with activity to chronic limb-threatening ischemia with risk of eventual tissue loss. Despite the significant global prevalence and debilitating consequences of the disease, non-option patients remain largely undertreated. Recognizing that ECs are well-known for their therapeutic role in enhancing perfusion and neovascularization in mouse models of critical limb ischemia, we hypothesize that a synergy of ECs with SMCs may achieve better recovery of perfusion and neovascularization than ECs or SMCs alone. In this study, we compared the therapeutic efficacy of hiPSC-ECs, hiPSC-SMCs, and hiPSC-ECs + hiPSC-SMCs for the treatment of critical limb disease using a NOD-SCID mouse model of hind limb ischemia (HLI). A total of 1 × 10⁶ hiPSC-ECs, 1 × 10⁶ hiPSC-SMCs, or 0.5 × 10⁶ hiPSC-ECs + 0.5 × 10⁶ hiPSC-SMCs were intramuscularly injected into the ischemic limb of the mouse on day 3 after HLI induction. Laser Doppler was recorded on day 0 before, days 3 (i.e., before cell administration) and 31 after HLI induction. Compared with only basal medium injection, hiPSC-ECs alone most effectively improved perfusion (63.8% after normalized non-ischemic limb, p < 0.001) and neovascularization (total vessel density = 180.9/magnification, p < 0.001) followed by co-administration of hiPSC-ECs + hiPSC-SMCs (perfusion = 59%, p = 0.008 and total vessel density = 170.5, p = 0.002) compared with basal medium injection (perfusion = 39.7% and total vessel density = 107.2), while hiPSC-SMCs administration achieved mild to moderate improvement (perfusion = 48.9% and total vessel density = 159.5, p = 0.01). When profiles of secreted growth factors or cytokines were determined using a protein array, 34 growth factors or cytokines involved in angiogenesis were highly enriched (at least >50% increase) in a hiPSC-ECs conditioned medium, including basic fibroblast growth factor and vascular endothelial growth factor, while there were 14 in hiPSC-SMCs conditioned medium. In conclusion, our results demonstrate that hiPSC-ECs alone most effectively improved perfusion and neovascularization, followed by co-administration of hiPSC-ECs + hiPSC-SMCs, which may be due to the abundance of growth factors and cytokines secreted by hiPSC-ECs. Future studies are warranted to evaluate the therapeutic efficacy of hiPSC-EC and hiPSC-SMC co-transplantation in aged animals with metabolic and cardiovascular comorbidities.

Developed micro-nano-materials have shown great promise to translate from bench to bed side, because they could be customized to bypass the physiological and pathological barriers, increase targeting efficiency, minimize off-target effects, optimize pharmacokinetics (PK) and pharmacodynamics (PD), decrease the therapeutic threshold dosage and side-effects, and ultimately boost the treatment outcomes and prognosis. [5,6] In this special issue, both reviews and original research articles are collected with the focused topic.So far, cancer is still one of the most life-threatening diseases for human beings worldwide. Although a great number of treatment paradigms and drugs have been invented, patients bearing cancer often suffer from severe side effects and a poor prognosis. For efficiently diagnosing cancer, the fluorescence imaging in the second near-infrared window (NIR-II) offers advantages such as deep tissue penetration, reduced autofluorescence, high spatiotemporal resolution, and excellent signal-to-background ratio, making it a powerful tool for diagnosis and imaging-guided drug delivery (IGDD) in cancer therapy. Roy et al. highlighted recent advancements in NIR-II IGDD systems, focusing on strategies to enhance drug loading, controlled release, and overcoming biological barriers. It categorizes these systems based on design elements, including intrinsic or extrinsic fluorophores and responsive drug delivery mechanisms. Moreover, they discussed the applications of NIR-II IGDD in various treatment modalities, such as chemotherapy, photodynamic, photothermal, chemodynamic, immunotherapy, gas therapy, and ion channel-targeted therapy, while also emphasizing multifunctional capabilities like real-time monitoring of drug release and therapeutic effects. [7] Isaí et al. used Coffea arabica green bean extract, doxorubicin, and commercial organic ligands (MUA) to synthesize gold nanoparticles through a green synthetic route. The nanoparticles showed good stability with a diameter of 40 nm at a physiological pH environment, but triggered release at tumoral pH microenvironment, and exhibited highly efficient tumor specificity and cytotoxicity to H69 lung cancer cells. This novel approach provides a promising avenue for cancer therapy by harnessing the distinct advantages of each component to enhance treatment effectiveness while minimizing adverse side effects. [8] Nowadays, with the increasing global aging population, osteoarthritis (OA) with closely related to aging and obesity, has become a representative degenerative disease and leads to an economic burden to society.The research in OA treatment based on nanomedicines has grown rapidly. The research direction generally is to customize nanocarriers to efficiently deliver therapeutic agents like drugs and nucleic acids to the pathological lesion of OA, in which the microenvironment is weakly acidic and full of synovial fluid. For example, hybrid DDS platforms combining hydrogels, liposomes, and particle-based carriers are being designed to localize within the synovial joint and alleviate pain and inflammation. Meanwhile, cartilagetargeting systems are being engineered to bind specific intra-cartilage components such as aggrecans, collagen type II, and chondrocytes, improving drug access to cellular and intracellular targets. These strategies aim to enable effective delivery of disease-modifying OA drugs, including emerging gene therapies. Recent clinical progress includes the approval of steroid-loaded polymeric microparticles for prolonged pain management. In preclinical studies, electrically charged biomaterials have demonstrated improved cartilage targeting and therapeutic outcomes. Ongoing clinical trials assessing viral vector-based gene therapies hold promise for durable, long-term treatment solutions for OA. [9] Shuai et al. updated the recent progress of pH-responsive nanomedicines for OA therapy, including the composition, synthetic routes, pH-responsive mechanism, targeting capability, and application examples. Special focus is on how to achieve targeting to the acidic microenvironment of the inflamed joints of OA. This work also emphasizes the integration of multiple stimuli-responsive mechanisms, such as pH, ultrasound, magnetic fields, and MMPs, to overcome the limitations of burst release and poor targeting seen in conventional DDS. A significant innovation lies in combining therapy and imaging functionalities, enabling real-time monitoring of disease progression. The paper also highlights underexplored targets like synovium and subchondral bone, encouraging a shift beyond cartilage-centric strategies. Moreover, it introduces advanced dual-drug delivery platforms capable of co-loading hydrophilic and hydrophobic drugs, achieving synergistic antiinflammatory, antioxidant, and cartilage-repair effects. The inclusion of multifunctional materials like circular brush zwitterionic polymers with joint lubrication and ROS-scavenging properties further enhances therapeutic performance. Finally, the work underlines the need to tailor nanoparticle design to OA's complex microenvironment, promoting the development of highly specific, biocompatible, and efficient nanocarriers for precise and multimodal OA therapy. [10] Recently, ionic liquid (IL)-based carriers as a new drug delivery platform have gained increasing attention in the pharmaceutical industry, owing to their merits including versatile chemical structures, ease in synthesis, good stability, and amphiphilic nature with efficient solubility to both hydrophilic and hydrophobic drugs. For example, the ILs like organic salts with melting points below 100 °C can serve as solvents, excipients, or active pharmaceutical ingredients (API-ILs), with some API-ILs combining drug molecules with biologically meaningful ions to improve pharmacokinetics and minimize polymorphism issues. [11] Ankit et al. offered a timely summary of IL-based carriers for drug delivery...

It remains a significant challenge to reactivate the cell cycle activity of adult mammalian cardiomyocytes (CMs). This study created a hypo-immunogenic human induced pluripotent stem cell (hiPSC) line using clustered regularly interspaced palindromic repeats (CRISPR)/Cas9 gene editing to knockout β2-microglobulin in hiPSCs (^B2MKOhiPSCs) for manufacturing nanovesicles (^B2MKOhiPSC-NVs). Approximately 9500 ^B2MKOhiPSC-NVs were produced from a single ^B2MKOhiPSC. Proteomic analyses indicated that, compared to ^B2MKOhiPSCs, the cargos of ^B2MKOhiPSC-NVs were enriched in spindle and chromosomal proteins, as well as proteins that regulate the cell cycle and scavenge reactive oxygen species (ROS). When administrated to hiPSCs derived CMs (hiPSC-CMs), ^B2MKOhiPSC-NVs reduced lactate dehydrogenase leakage and apoptosis in hypoxia-cultured hiPSC-CMs through activating the AKT pathway, protected hiPSC-CMs from H₂O₂-induced damage by ROS scavengers in the NV cargo, increased hiPSC-CM proliferation via the YAP pathway, and were hypoimmunogenic when co-cultured with human CD8⁺ T cells or delivered to mice. Furthermore, when ^B2MKOhiPSC-NVs or 0.9 % NaCl were intramyocardially injected into mice after cardiac ischemia/reperfusion injury, cardiac function and infarct size, assessed 4 weeks later, were significantly improved in the ^B2MKOhiPSC-NV group, with increased mouse CM survival and cell cycle activity. Thus, the proteins in the ^B2MKOhiPSC-NV cargos convergently activated the AKT pathway, scavenged ROS to protect CMs, and upregulated YAP signaling to induce CM cell cycle activity. Thus, ^B2MKOhiPSC-NVs hold great potential for cardiac protection and regeneration.

Stem cell biology, and developmental biology are transforming the way we understand and treat heart failure.In this Special Issue, entitled "Translational Advances in Cardiovascular Therapy:From Bench to Bedside", six original research papers depict some underlying mechanisms precipitating in cardiac disorders and modulatory measures to mitigate the dysfunction of the myocardium. The studies include an understanding of zinc oxide nanoparticles (ZnO NPs) in induction of arrhythmia, how monocyte adhesion to and transmigration through endothelium, immunomodulation role of CD16 + monocyte in myocardial infarction, and the association between iron metabolism, transferrin saturation level, and stress hyperglycemia ratio with cardiac diseases.Liu et al., [1] explored the effect and underlying mechanism of ZnO NPs exposure on cardiac function, especially during acute exposure. The study showed that acute exposure to ZnO NPs markedly decreased voltage-gated sodium current (INa) and long-lasting calcium current (Ica-L), resulting in a reduced amplitude and shortened action potential duration in cardiomyocytes. These changes not only prolonged PR-interval and blocked A-V conduction that triggered cardiac arrhythmia, but also led to a diminished Large animal models, such as pigs, are essential for translational cardiovascular research as they more closely resemble human anatomy, physiology, and function.Ascione et al., [3] investigated the feasibility, safety and efficacy of hCD16 + monocytes for the treatment of myocardial infarction (MI) in a pig model. Cardiac magnetic resonance analysis suggested that although LVEF did not differ significantly between groups, CD16 + monocyte administration resulted in 14.5 g scar reduction (from 25.45 ± 8.24 to 10.8 ± 3.4 gr; -55%) as compared to 6.4 g scar reduction (from 18.83 ± 5.06 to 12.4 ± 3.9gr; -30%) in the control group 30 days after treatment. In addition, higher tissue levels of neo-angiogenesis, myofibroblast and IL-6 and lower levels of TGF-β were observed in the hCD16 + monocytes treated group. These data demonstrate that the use of hCD16 + monocytes in acute MI is feasible, safe, and associated with reduced LV scar size, increased neo-angiogenesis, myofibroblasts, and IL-6.In addition to basic and translation studies, three original articles reported clinical findings. Zhu et al., [4] identified eight differentially expressed genes related to iron metabolism. These genes are mainly involved in the cellular stress response. A logistic regression model based on these genes achieved an AUC of 0.64-0.65 in the diagnosis of coronary heart disease, indicating these genes may have diagnostic potential for coronary heart disease.Wang et al., [5] examined the data from the USA National Health and Nutrition Examination Survey (NHANES, 2017-2020.03) in adults aged ≥40 years and explored the association between serum transferrin saturation levels and heart failure. They found that participants with heart failure had significantly lower serum transferrin saturation levels compared to those without heart failure. After fully adjusting for potential confounders, weighted multiple logistic regression models revealed a 2.6% reduced in the risk of heart failure when each unit of serum transferrin saturation level increased.These findings suggest a negative association between serum transferrin saturation levels and heart failure among middle-aged and older adults in the United States.Liu et al., [6] assessed the value of stress hyperglycemia ratio in determining outcomes in patients with atrial fibrillation in intensive care unit. Among patients with critical atrial fibrillation, those with the highest stress hyperglycemia ratio quartiles exhibited an increased risk of 365-day all-cause mortality (HR:1.32, 95%CI=1.06-1.65).Notably, in subgroup analyses, the prognostic value of atrial fibrillation was particularly pronounced in patients with comorbid condition, hypertension. This highlights a positive association between stress hyperglycemia ratio caused by acute glycemic dysregulation and all-cause mortality in critically ill patients with atrial fibrillation. In summary, this Special Issue provides an additional facet to understanding causes of cardiac disorders, as well as put forth potential therapeutic interventions targeting the dysfunctional myocardium. Ensuing research is crucial to close the gap between existing knowledge with the mechanisms highlighted in this Special Issue as well as further validate the diagnostic and prognostic tools described.

BACKGROUND: When human induced pluripotent stem cells (hiPSCs) that CCND2-OE (overexpressed cyclin-D2) were differentiated into cardiomyocytes ( ^CCND2-OE hiPSC-CMs) and administered to the infarcted hearts of immunodeficient mice, the cells proliferated after administration and repopulated >50% of the scar. Here, we knocked out human leukocyte antigen class I and class II expression in ^CCND2-OE hiPSC-CMs ( ^KO/OE hiPSC-CMs) to reduce the cells’ immunogenicity and then assessed the therapeutic efficacy of ^KO/OE hiPSC-CMs for the treatment of myocardial infarction. METHODS: ^KO/OE hiPSC-CM and wild-type hiPSC-CM ( ^WT hiPSC-CM) spheroids were differentiated in shaking flasks, purified, characterized, and intramyocardially injected into pigs after ischemia/reperfusion injury; control animals were injected with basal medium. Cardiac function was evaluated via cardiac magnetic resonance imaging, and cardiomyocyte proliferation was assessed via immunostaining and single-nucleus RNA sequencing. RESULTS: Measurements of cardiac function and scar size were significantly better in pigs treated with ^KO/OE hiPSC-CM spheroids than in animals treated with medium or ^WT hiPSC-CM spheroids. ^KO/OE hiPSC-CMs were detected for just 1 week after administration, but assessments of cell cycle activity and proliferation were significantly higher in the endogenous pig cardiomyocytes of the hearts from the ^KO/OE hiPSC-CM spheroid group than in those from the other 2 groups. Single-nucleus RNA-sequencing analysis identified a cluster of proliferating cardiomyocytes that was significantly more prevalent in the ^KO/OE hiPSC-CM spheroid–treated hearts (3.65%) than in the hearts from the medium (0.89%) or ^WT hiPSC-CM spheroid (1.33%) groups at week 1. YAP (Yes-associated protein) protein levels and nuclear localization were also significantly upregulated in pig cardiomyocytes after treatment with ^KO/OE hiPSC-CM spheroids. Follistatin, which interacts with the HIPPO/YAP pathway, was significantly more abundant in the medium from ^KO/OE hiPSC-CM spheroids than ^WT hiPSC-CM spheroids (30.29±2.39 versus 16.62±0.83 ng/mL, P =0.0056). Treatment with follistatin increased ^WT hiPSC-CM cell counts by 28.3% over 16 days in culture and promoted cardiomyocyte proliferation in the infarcted hearts of adult mice. CONCLUSIONS: ^KO/OE hiPSC-CM spheroids significantly improved cardiac function and reduced infarct size in pig hearts after ischemia/reperfusion injury by secreting follistatin, which upregulated HIPPO/YAP signaling and proliferation in endogenous pig cardiomyocytes.

Chronic ischemic limb disease often leads to amputation, which remains a significant clinical problem. Smooth-muscle cells (SMCs) are crucially involved in the development and progression of many cardiovascular diseases, but studies with primary human SMCs have been limited by a lack of availability. Here, we evaluated the efficiency of two novel protocols for differentiating human induced-pluripotent stem cells (hiPSCs) into SMCs and assessed their potency for the treatment of ischemic limb disease. hiPSCs were differentiated into SMCs via a conventional two-dimensional (2D) protocol that was conducted entirely with cell monolayers, or via two protocols that consisted of an initial five-day three-dimensional (3D) spheroid phase followed by a six-day 2D monolayer phase (3D + 2D differentiation). The 3D phases were conducted in shaker flasks on an orbital shaker (the 3D + 2D shaker protocol) or in a PBS bioreactor (the 3D + 2D bioreactor protocol). Differentiation efficiency was evaluated via the expression of SMC markers (smooth-muscle actin [SMA], smooth muscle protein 22 [SM22], and Calponin-1), and the biological activity of the differentiated hiPSC-SMCs was evaluated via in-vitro assessments of migration (scratch assay), contraction in response to the treatment with a prostaglandin H2 analog (U46619), and tube formation on Geltrex, as well as in-vivo measurements of perfusion (fluorescence angiography) and vessel density in the limbs of mice that were treated with hiPSC-SMCs after experimentally induced hind-limb ischemia (HLI). Both 3D + 2D protocols yielded > 5.6 × 107 hiPSC-SMCs/differentiation, which was ~ nine-fold more than that produced via 2D differentiation, and flow cytometry analyses confirmed that > 98% of the 3D + 2D-differentiated hiPSC-SMCs expressed SMA, > 81% expressed SM22, and > 89% expressed Calponin-1. hiPSC-SMCs obtained via the 3D + 2D shaker protocol also displayed typical SMC-like migratory, contraction, and tube-formation activity in-vitro and significantly improved measurements of perfusion, vessel density, and SMA-positive arterial density in the ischemic limb of mouse HLI model. Our dynamic 3D + 2D protocols produced an exceptionally high yield of hiPSC-SMCs. Transplantation of these hiPSC-SMCs results in significantly improved recovery of ischemic limb after ischemic injury in mice.